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Abstract:

An arrangement is provided in at least one embodiment, having a magazine
for supplying a plurality of microfluidic devices. The microfluidic
devices each contain at least one element/device for binding at least one
biological molecule, wherein the at least one element/device for binding
the at least one biological molecule can be moved relative to the
microfluidic device. A sample presumably containing biological molecules
to be examined is introduced into the microfluidic device. The biological
molecule to be examined is bound by the at least one element/device for
binding the biological molecule. In at least one embodiment, the at least
one element/device for binding the at least one biological molecule, or
the substrate-molecule complex, can then be moved in the microfluidic
device, e.g. in accordance with a predetermined reaction sequence, for
example by means of a magnetic field. The microfluidic device is
transported through the arrangement.

Claims:

1. An arrangement for processing a plurality of samples for analysis, the
arrangement comprising: a receptacle for a microfluidic device, including
means for binding at least one biological molecule provided in the
microfluidic device; means for moving the microfluidic device in the
arrangement along at least one direction of movement; and at least one
magazine for supplying a plurality of microfluidic devices.

2. The arrangement as claimed in claim 1, further comprising: a unit for
moving means, provided in the microfluidic device, for binding at least
one biological molecule relative to the microfluidic device.

3. The arrangement as claimed in claim 2, wherein the unit for moving
means provided in the microfluidic device for binding at least one
biological molecule relative to the microfluidic device comprises a
magnetic field generator.

4. The arrangement as claimed in claim 1, further comprising: means for
amplifying the biological molecule in the microfluidic device.

5. The arrangement as claimed in claim 1, further comprising: means for
detecting the biological molecule.

6. The arrangement as claimed in claim 1, further comprising: a unit for
introducing a sample into a microfluidic device.

7. The arrangement as claimed in claim 1, further comprising: a container
for collecting used microfluidic devices.

8. The arrangement as claimed in claim 1, wherein the at least one
magazine is a stack magazine, in which the microfluidic devices are
stackable.

9. The arrangement as claimed in claim 1, wherein the at least one
magazine is a drum magazine, in which the microfluidic devices are
rollable on a roll.

10. The arrangement as claimed in claim 1, further comprising: means for
detecting a coding of a microfluidic device.

11. A method for processing a plurality of samples for analysis, the
method comprising: a) supplying an arrangement with a plurality of
microfluidic devices, wherein the arrangement has at least one magazine
for supply with microfluidic devices and wherein the microfluidic devices
each contain at least one element for binding at least one biological
molecule; b) introducing a first sample, containing at least one
biological molecule to be examined, into one of the microfluidic devices;
c) binding the biological molecule to be examined by the at least one
element for binding at least one biological molecule; and d) repeating
steps b)-c) with the further samples until all the samples to be
processed have been processed, wherein the microfluidic device with the
introduced sample is moved in the arrangement along at least one
direction of movement.

12. The method as claimed in claim 11, wherein the at least one element
means for binding the at least one biological molecule is movable
relative to the microfluidic device.

13. The method as claimed in claim 11, wherein the introduction of the
samples, containing biological molecules to be examined, into the
respective microfluidic devices is effected before the supply of the
arrangement with microfluidic devices.

14. The method as claimed in claim 11, wherein the at least one element
for binding the at least one biological molecule have a substrate that
can be linked to the molecule to form a substrate-molecule complex.

15. The method as claimed in claim 14, wherein, after the binding of the
molecule to the substrate, the substrate-molecule complex is separated
from the rest of the sample.

16. The method as claimed in claim 15, wherein the separation of the
substrate-molecule complex from the rest of the sample is effected by
moving the substrate-molecule complex relative to the rest of the sample.

17. The method as claimed in claim 11, wherein the at least one element
for binding the at least one biological molecule comprise at least one
magnetic element.

18. The method as claimed in claim 17, wherein a magnetic field is used
for moving the at least one element for binding the at least one
biological molecule relative to the microfluidic device.

19. The method as claimed in claim 11, further comprising: amplifying the
biological molecule by way of an amplification reaction.

20. The method as claimed in claim 11, further comprising: detecting the
biological molecule.

21. The method as claimed in claim 11, wherein the at least one element
for binding the at least one molecule is moved along a reaction section
in the microfluidic device, which leads into at least one process
chamber.

22. The method as claimed in claim 21, wherein the at least one element
for binding the at least one molecule is moved along a reaction section
in the microfluidic device through a plurality of process chambers.

23. The method as claimed in claim 21, wherein the reaction section is
oriented essentially along the at least one direction of movement in the
arrangement.

24. The method as claimed in claim 21, wherein, in the microfluidic
device, a plurality of samples are processed simultaneously in a
corresponding number of reaction sections which are arranged essentially
parallel in the microfluidic device.

25.-28. (canceled)

29. The method as claimed in claim 12, wherein the introduction of the
samples, containing biological molecules to be examined, into the
respective microfluidic devices is effected before the supply of the
arrangement with microfluidic devices.

30. The method as claimed in claim 12, wherein the at least one element
for binding the at least one biological molecule have a substrate that
can be linked to the molecule to form a substrate-molecule complex.

31. The method as claimed in claim 22, wherein the reaction section is
oriented essentially along the at least one direction of movement in the
arrangement.

32. An arrangement for processing a plurality of samples for analysis,
the arrangement comprising: a receptacle for a microfluidic device,
including at least one element to bind at least one biological molecule
provided in the microfluidic device; at least one device to move the
microfluidic device in the arrangement along at least one direction of
movement; and at least one magazine to supply a plurality of microfluidic
devices.

33. The arrangement as claimed in claim 32, further comprising: a unit
for moving device, provided in the microfluidic device, to bind at least
one biological molecule relative to the microfluidic device.

34. The arrangement as claimed in claim 32, further comprising: at least
one device to amplify the biological molecule in the microfluidic device.

35. The arrangement as claimed in claim 32, further comprising: at least
one device to detect the biological molecule.

36. The arrangement as claimed in claim 32, wherein the at least one
magazine is a stack magazine, in which the microfluidic devices are
stackable.

37. The arrangement as claimed in claim 32, wherein the at least one
magazine is a drum magazine, in which the microfluidic devices are
rollable on a roll.

Description:

PRIORITY STATEMENT

[0001] This application is the national phase under 35 U.S.C. §371 of
PCT International Application No. PCT/EP2007/062977 which has an
International filing date of Nov. 29, 2007, which designated the United
States of America, and which claims priority on German patent application
number DE 10 2006 057 300.5 filed Dec. 5, 2006, the entire contents of
each of which are hereby incorporated herein by reference.

FIELD

[0002] At least one embodiment of the invention generally relates to an
arrangement for processing a plurality of samples for analysis, which has
an arrangement, microfluidic devices for receiving samples and at least
one device for moving the microfluidic devices in the arrangement. At
least one embodiment of the invention furthermore relates to a method for
processing a plurality of samples for analysis.

BACKGROUND

[0003] In biotechnological analysis, in recent years high throughput
methods (high throughput screening, HTS) have been developed in order to
be able to process a large number of samples in a short time. Hole plate
formats have predominantly been used here, e.g. 96-hole plates or
384-hole plates, wherein each hole or each depression in a plate
constitutes a reaction vessel. The disadvantage of such methods is that
liquids have to be pipetted over from supply vessels to the plate or from
plate to plate, which is mechanically complicated and entails risks of
contamination.

[0004] In addition, fully integrated microfluidic analysis devices have
also been developed, wherein, instead of reaction vessels, process
chambers are used which are connected via lines or channels, as described
e.g. in DE 101 11 457 A1. These devices can be contained in a fully
encapsulated manner in a cartridge, a card-like flat structure, wherein
process chambers for sample processing, amplification of analytes, e.g.
nucleic acids, and for detection of analytes, e.g. in the form of
biochips with nucleic acid microarrays, are provided in the device.
Analysis devices of this type have the advantage that the analysis can
proceed completely in the encapsulated analysis device, such that risks
of contamination or operating errors are largely precluded.

[0005] Devices of this type can be used for analyzing nucleic acids, e.g.
DNA sequences or RNA sequences, proteins and other biomolecules. Even
complex assay sequences can be carried out in a manner free of
contamination and errors in such an analysis device in microfluidic
arrangements of process chambers and connecting channels. One
disadvantage of these systems, however, is the low sample throughput,
that is to say the small number of assays that can be carried out per
time. Particularly in the case of nucleic acid-based systems that require
amplification of the DNA or RNA, a total duration of the assay of one
hour or more is by no means an exception. Generally, in this case firstly
the sample is introduced manually into the analysis device, and the
latter is then inserted into a control or read-out unit, in which the
process steps are processed automatically. At the end of the assay, the
analysis device is manually removed from the control unit. This process
sequence requires regular manual intervention by the operating personnel
and significantly restricts the throughput.

[0006] Theoretically, it is possible to use fully integrated diagnostic
systems for complex assays with many biological issues (e.g.
multiparameter studies such as the cytochrome P 450 analysis or CFTR) in
central laboratories as well. In this case, however, the low throughput
is a major disadvantage and leads to prohibitively high costs. In
established high throughput methods, e.g. the above-described methods on
hole plate formats, the processing of the samples for analysis is
complicated. This processing can be carried out with comparatively little
complexity in microfluidic devices, however, e.g. by disrupting the
sample, binding the analytes to magnetic substrates, so-called magnetic
beads, fixing the substrate-analyte complex by way of an external
magnetic field in the analysis device and removing undesirable sample
constituents by rinsing the fixed substrate-analyte complexes with a
washing liquid, as described e.g. in DE 101 11 520 B4. However, methods
of this type have not had high throughput capability heretofore.

SUMMARY

[0007] At least one embodiment of the present invention provides an
arrangement and a method for processing a plurality of samples for
analysis which can be implemented in a fully integrated analysis device
and is simultaneously suitable for processing high numbers of samples.

[0008] The arrangement according to at least one embodiment of the
invention is for processing a plurality of samples for analysis
comprising: [0009] a) a receptacle for a microfluidic device, wherein
at least one device/element for binding at least one biological molecule
is provided in the microfluidic device; and [0010] b) at least one
device/element for moving the microfluidic device in the arrangement
along at least one predetermined direction of movement; [0011] wherein
the arrangement has at least one magazine for supplying a plurality of
microfluidic devices.

[0012] Preferably, the arrangement has a unit for moving provided in the
microfluidic device for binding at least one biological molecule relative
to the microfluidic device. The unit preferably comprises a magnetic
field generator.

[0013] The expression "microfluidic device" relates to a device in which
fluid volumes in the microliters range can be manipulated, e.g.
microfluidic cartridges such as are generally known in the art.

[0014] Preferably, the arrangement according to at least one embodiment of
the invention furthermore has at least one device/element for amplifying
the biological molecule in the microfluidic device.

[0015] Furthermore, the arrangement according to at least one embodiment
of the invention comprises at least one device/element for detecting the
biological molecule.

[0016] The arrangement according to at least one embodiment of the
invention furthermore preferably comprises a unit for introducing a
sample into a microfluidic device.

[0017] In accordance with one preferred aspect of at least one embodiment
of the invention, the arrangement comprises a container for collecting
used microfluidic devices.

[0018] In accordance with one preferred aspect of at least one embodiment
of the invention, the arrangement comprises a stack-like magazine in
which the microfluidic devices can be stacked.

[0019] In accordance with an alternative aspect of at least one embodiment
of the invention, the arrangement comprises a drum-like magazine in which
the microfluidic devices can be rolled up on a roll.

[0020] In accordance with a further preferred aspect of at least one
embodiment of the invention, the arrangement comprises at least one
device/element for detecting a coding of a microfluidic device.

[0021] According to at least one embodiment of the invention, a sample
that presumably contains biological molecules to be examined is
introduced into the microfluidic device, wherein, in the microfluidic
device, the biological molecule to be examined is bound by the at least
one device/element for binding and processing of the sample is thus made
possible. A further microfluidic device can be loaded from the magazine,
and can be filled with the next sample. As an alternative, a plurality of
microfluidic devices can be filled with the samples before being
introduced into the arrangement. The microfluidic devices are transported
through the arrangement along the at least one predetermined direction of
movement and the samples can in this way be processed successively in an
automated manner.

[0022] Preferably, the at least one device/element for binding the at
least one biological molecule are embodied as a substrate that can be
linked to the biological molecule to be examined to form a
substrate-molecule complex.

[0023] In accordance with one aspect of at least one embodiment of the
present invention, the substrate has a protein-binding property, which
can preferably be embodied as an antibody directed to the biological
molecule.

[0024] In accordance with an alternative aspect of at least one embodiment
of the present invention, the substrate has a nucleic acid-binding
property, wherein the nucleic acid-binding property is preferably
embodied non-sequence-specifically, e.g. as silane, or as probe
oligonucleotid (sequence-specifically).

[0025] In accordance with a further aspect of at least one embodiment of
the present invention, the substrate has both at least one
protein-binding property and at least one nucleic acid-binding property.

[0026] Preferably, the at least one device/element for binding at least
one biological molecule comprise at least one magnetic element, e.g.
magnetic bead, which can be moved and/or fixed by a magnetic field.

[0027] Preferably, the arrangement has at least one device/element for
amplifying the molecule. This can comprise, e.g. if the molecule is a
nucleic acid, an amplification chamber in the microfluidic device, in
which an amplification reaction, e.g. the polymerase chain reaction
(PCR), or a comparable amplification method, can take place. Heating
and/or cooling elements, e.g. peltier elements, can be provided in the
arrangement in order to carry out such a reaction.

[0028] Furthermore, the arrangement according to at least one embodiment
of the invention preferably has at least one device/element for detecting
the molecule. The detection can be effected e.g. by magnetic, optical,
florescence-optical, electrochemical, gravimetric and other suitable
methods. For this purpose, a detection chamber is provided in the
microfluidic device, which detection chamber can have a nucleic acid
microarray, for example, on which probe oligonucleotides for the
detection of nucleic acid molecules are provided. Electrochemical
detection is particularly preferred. For this purpose, an electrochemical
sensor, e.g. in the form of electrodes, is provided in the microfluidic
device. At least one device/element for measuring currents and/or
voltages is/are provided in the arrangement. A corresponding measurement
method that can be used in this case is described e.g. in DE 101 26 341
A1. In accordance with an alternative aspect, magnetic detection is
preferred. For this purpose, a magnetoresistive sensor can be provided in
the arrangement.

[0029] In accordance with a further aspect of at least one embodiment of
the present invention, the microfluidic device comprises at least one
process chamber which at least temporarily contains the at least one
device/element for binding at least one biological molecule. The at least
one process chamber can be embodied as a processing chamber for using the
at least one device/element for binding the at least one biological
molecule, as an amplification chamber for using the at least one
device/element for amplifying the at least one biological molecule,
and/or as a detection chamber for detecting the at least one biological
molecule. Provision can preferably be made of a plurality of process
chambers which are arranged along a reaction section and can be connected
by lines at least occasionally. The lines can be embodied as microfluidic
channels with valves fitted thereto. The valves can be embodied as simple
elastic pinch valves or magnetically controllable valves in order to
fluidically separate the different process chambers from one another. The
valves can also be embodied in other ways known to the person skilled in
the art.

[0030] In accordance with one preferred aspect of at least one embodiment
of the present invention, a plurality of groups of process chambers can
be provided in a microfluidic device, wherein the process chambers in a
group are preferably arranged in each case along a reaction section and
the process chambers of a group along the respective reaction section can
be fluidically connected by lines at least occasionally. In this way it
is possible to realize a plurality of sample sections e.g. in a parallel
fashion on the microfluidic device. Sample ports arranged parallel can be
situated at one end of the respective reaction sections, which ports can
be sealed by septa. At the other end, there can be provided as detection
devices/elements e.g. correspondingly a plurality of microarrays arranged
parallel or else alternatively a microarray common to the individual
reaction sections and serving for detecting the target molecules in all
the biological samples applied. The sample ports can be connected to the
microarrays along the reaction section via various process chambers (e.g.
processing, washing and amplification chambers) and lines.

[0031] A microfluidic device of this type can be used as a single-use
element in the arrangement according to the invention. The single-use
element can be embodied as an elongate device, e.g. in the form of a
cartridge, that is to say a card-like flat structure. Preferably, the
chambers and lines are oriented along the reaction section in the
elongate device along the at least one direction of movement with which
the elongate device is transported through the control unit. It is noted
that the microfluidic device having a plurality of parallel reaction
sections as described in this paragraph is considered to constitute an
autonomous embodiment of the invention which is independent of the rest
of the arrangement and which likewise achieves the object formulated
initially.

[0032] Furthermore, a container for collecting the used microfluidic
devices is preferably provided in the arrangement.

[0033] The microfluidic devices can be discarded and disposed of after
single use. However, it is also conceivable that they can be reused, e.g.
after cleaning or regeneration.

[0034] The arrangement can have a stack-like magazine in which the
microfluidic devices are stacked. As an alternative, it is also possible
to provide a drum-like magazine, for example, in which the microfluidic
devices are rolled up on a roll.

[0035] Preferably, the arrangement has a unit for introducing a sample
into a microfluidic device. This can be configured as an automated
pipetting device, for example, which can pipette a sample into the
microfluidic device. If a plurality of parallel reaction sections for the
parallel processing of samples are provided on the microfluidic device,
the unit for introducing a sample into the microfluidic device preferably
has a corresponding number of channels in order to introduce the
corresponding number of samples in one work step.

[0036] At least one device/element for moving or transporting the
microfluidic device along at least one predetermined direction of
movement is/are provided in the control unit; these transport
devices/elements can be embodied e.g. in the form of a conveyor belt.

[0037] In accordance with a further aspect of at least one embodiment of
the present invention, at least one device/element for moving the
substrate-molecule complex relative to the microfluidic device are
furthermore provided, which preferably comprise a magnetic field
generator. By moving the microfluidic device relative to the magnetic
field generator, or by moving the magnetic field generator relative to
the microfluidic device, it is possible for the substrate-molecule
complex having magnetic beads to be moved relative to the microfluidic
device, that is to say e.g. along the reaction path through the process
chambers. In this case, the magnetic beads are retained in the magnetic
field of the magnetic field generator, while the microfluidic device is
moved relative to the magnetic field generator (or vice versa).

[0038] In accordance with a further aspect of at least one embodiment of
the present invention, the microfluidic devices have a marking by which
they can be coded. In this way it is possible to detect an assignment
between applied sample and the single-use element. For this purpose, at
least one device/element for detecting the marking are preferably
provided in the arrangement. The marking can comprise a conventional type
of marking known to the person skilled in the art, e.g. a bar code, an
RFID, or the like. Corresponding devices/elements for reading out the
marking are then preferably present in the arrangement. Furthermore, the
arrangement can be connected via interfaces to a data processing system
that is used to register the microfluidic devices on the basis of the
marking and to store data read out. Preferably, microfluidic devices once
used can be rendered invalid by way of the data processing system, in
order to preclude multiple reading.

[0039] The following procedure is performed when processing a plurality of
samples for analysis:

[0040] An arrangement is provided which has a magazine for the supply of a
plurality of microfluidic devices. The microfluidic devices each contain
at least one device/element for binding at least one biological molecule,
wherein the at least one device/element for binding the at least one
biological molecule can be moved relative to the microfluidic device.

[0041] A sample that presumably contains biological molecules to be
examined is introduced into the microfluidic device. Optionally, the
sample can firstly be disrupted in the microfluidic device, e.g. by using
a lysis buffer. The biological molecule to be examined is bound by the at
least one device/element for binding the biological molecule. Preferably,
the at least one device/element for binding the at least one biological
molecule are embodied as a substrate that can be linked to the molecule
to form a substrate-molecule complex. The at least one device/element for
binding the at least one biological molecule, or the substrate-molecule
complex, can then be moved in the microfluidic device, e.g. in accordance
with a predetermined reaction sequence.

[0042] In accordance with a further aspect of at least one embodiment of
the present invention, after the binding of the molecule, the
substrate-molecule complex can be separated from the remainder of the
sample. This can be done by moving the substrate-molecule complex
relative to the rest of the sample volume, e.g. by magnetically fixing
the substrate-molecule complex and rinsing away the sample.

[0043] At least one device/element for pumping fluids into the
microfluidic device and/or out of the microfluidic device can be provided
in the arrangement. They can be embodied e.g. as lines, channels, with
corresponding filling or extracting units, using corresponding fluid
transport systems, e.g. piston pumps, peristaltic pumps and other pumps
known to the person skilled in the art.

[0044] In accordance with a further aspect of at least one embodiment of
the present invention, the molecule can also be separated from the
substrate again in the course of the method, e.g. by separating the
substrate-molecule complex bond, e.g. by heating, changing the salt
concentration or the like.

[0045] In accordance with one preferred aspect of at least one embodiment
of the invention, the method has an additional step of amplification of
the molecule by way of an amplification reaction. Furthermore, the method
preferably has the additional step of detection of the molecule.

[0046] In the method according to at least one embodiment of the
invention, the at least one device/element for binding the at least one
molecule is preferably moved along a reaction section in the microfluidic
device, which leads into at least one process chamber.

[0047] Preferably, a plurality of process chambers are arranged along the
reaction section. In accordance with a further aspect of at least one
embodiment of the present invention, in the method, in the microfluidic
device, a plurality of samples are processed simultaneously in a
corresponding number of reaction sections which are arranged essentially
parallel in the microfluidic device.

[0048] Preferably, the microfluidic devices are loaded from the magazine,
pass through the arrangement along the at least one predetermined
direction of movement and are then ejected from the arrangement or
transported into a container for collecting used microfluidic devices.

[0049] At least one embodiment of the invention furthermore relates to a
method for processing a plurality of samples for analysis, having the
following steps: [0050] a) supply of an arrangement with a plurality of
microfluidic devices, wherein the arrangement has at least one magazine
for supply with microfluidic devices and wherein the microfluidic devices
each contain at least one device/element for binding at least one
biological molecule; [0051] b) introduction of a first sample, containing
at least one biological molecule to be examined, into one of the
microfluidic devices; [0052] c) binding of the biological molecule to be
examined by the at least one device/element for binding at least one
biological molecule; and [0053] d) repetition of steps b)-c) with the
further samples until all the samples to be processed have been
processed; [0054] wherein the microfluidic device with the introduced
sample is moved in the arrangement along at least one predetermined
direction of movement.

[0055] In this case, the at least one device/element for binding the at
least one biological molecule can preferably be moved relative to the
microfluidic device.

[0056] Preferably, the introduction of the samples, containing biological
molecules to be examined, into the respective microfluidic devices is
effected before the supply of the arrangement with microfluidic devices.

[0057] In accordance with one preferred aspect of the method according to
at least one embodiment of the invention, the at least one device/element
for binding the at least one biological molecule are embodied as a
substrate that can be linked to the molecule to form a substrate-molecule
complex.

[0058] Preferably, after the binding of the molecule to the substrate, the
substrate-molecule complex is separated from the rest of the sample.

[0059] In accordance with one preferred aspect of at least one embodiment
of the method according to the invention, the separation of the
substrate-molecule complex from the rest of the sample is effected by
moving the substrate-molecule complex relative to the rest of the sample.

[0060] In accordance with one preferred aspect of at least one embodiment
of the method according to the invention, the at least one device/element
for binding the at least one biological molecule comprise at least one
magnetic element.

[0061] In accordance with one preferred aspect of the method according to
at least one embodiment of the invention, a magnetic field is used for
moving the at least one device/element for binding the at least one
biological molecule relative to the microfluidic device.

[0062] In accordance with one preferred aspect of the method according to
at least one embodiment of the invention, the method according to the
invention has the additional step of amplification of the biological
molecule by way of an amplification reaction.

[0063] In accordance with one preferred aspect of the method according to
at least one embodiment of the invention, the method according to at
least one embodiment of the invention has the additional step of
detection of the biological molecule.

[0064] In accordance with one preferred aspect of the method according to
at least one embodiment of the invention, the biological molecule is
detected magnetically, electrochemically or optically.

[0065] In accordance with one preferred aspect of the method according to
at least one embodiment of the invention, the at least one device/element
for binding the at least one molecule is moved along a reaction section
in the microfluidic device, which leads into at least one process
chamber.

[0066] In accordance with one preferred aspect of the method according to
at least one embodiment of the invention, the at least one device/element
for binding the at least one molecule is moved along a reaction section
in the microfluidic device through a plurality of process chambers.

[0067] In accordance with one preferred aspect of the method according to
at least one embodiment of the invention, the reaction section is
oriented essentially along the at least one predetermined direction of
movement in the arrangement.

[0068] In accordance with one preferred aspect of the method according to
at least one embodiment of the invention, in this case, in the at least
single-use element, a plurality of samples are processed simultaneously
in a corresponding number of reaction sections which are arranged
essentially parallel in the microfluidic device.

[0069] Preferably, the microfluidic devices are loaded from the magazine,
pass through the arrangement along the at least one predetermined
direction of movement and are then ejected from the arrangement or
transported into a container for collecting used microfluidic devices.

BRIEF DESCRIPTION OF THE DRAWINGS

[0070] Further aspects, properties and advantages of the present invention
are illustrated on the basis of the following description of example
embodiments and the appended drawings, in which:

[0071]FIG. 1 shows a schematic illustration of a first embodiment of a
microfluidic device for receiving a sample in the arrangement according
to an embodiment of the invention;

[0072]FIG. 2 shows a second embodiment of a microfluidic device in the
arrangement according to an embodiment of the invention;

[0073]FIG. 3 shows a third embodiment of the microfluidic device in the
arrangement according to an embodiment of the invention;

[0074]FIG. 4 shows a first embodiment of the arrangement according to an
embodiment of the invention in a first operating state;

[0075]FIG. 5 shows a first embodiment of the arrangement according to an
embodiment of the invention in a second operating state;

[0076]FIG. 6 shows a first embodiment of the arrangement according to an
embodiment of the invention in a third operating state; and

[0077]FIG. 7 shows a second embodiment of the arrangement according to an
embodiment of the invention.

DETAILED DESCRIPTION OF THE EXAMPLE EMBODIMENTS

[0078]FIG. 1 schematically illustrates a microfluidic device which is
used in the arrangement according to the invention, in the form of a
cartridge 1. The latter is embodied as a card-like flat structure and can
be produced e.g. as a plastic injection-molded part, depressions present
therein being configured in the form of chambers and channels. Reagents
required for the subsequent processes and reactions, e.g. in the form of
dry reagents, can be introduced into the cartridge, e.g. by being spotted
on in the corresponding chambers. Sealing the upwardly open cartridge,
e.g. with a plastic film, creates a closed microfluidic device with
process chambers and lines situated therein. The cartridge 1 comprises a
processing chamber 3, a disruption chamber 5, a washing chamber 7, an
amplification chamber 9 and a detection chamber 11. The processing
chamber 3 comprises a filling opening 13, via which the sample to be
examined can be introduced into the processing chamber 3 for example by
way of a syringe or pipette. The process chambers 5, 7, 9, 11 are
connected via a microchannel 15 to an opening 17, via which water or
buffer can be introduced into the process chambers in a manner known per
se.

[0079] The filling opening 13 and/or opening 17 can be closed off by way
of a septum in order to ensure sterility and/or to prevent contaminations
and entrainments. Apart from the processing chamber 3, each process
chamber 5, 7, 9, 11 has a venting opening 19 closed off by a
gas-permeable membrane, for example. It can thus be ensured that gas can
leave the process chambers, but liquid cannot leave them. A lysis reagent
31, e.g. in dry form, is stored beforehand in the processing chamber. The
lysis reagent is dissolved by the introduction of the (liquid) sample,
e.g. blood or some other sample liquid. Biological structures, e.g.
cells, bacteria, viruses, are lysed by the dissolved lysis reagent and
release biological molecules contained therein.

[0080] The sample is then displaced from the chamber 3 into the chamber 5
via the line 23, e.g. by subsequent rinsing with buffer. Magnetic beads
21 in the dry state are stored beforehand in the chamber 5, and, as a
result of the sample being transferred into the chamber 5, the magnetic
beads 21 are suspended and distributed in the sample. Probe
oligonucleotides are provided on the magnetic beads, and bind target
molecules sought, e.g. nucleic acids complementary to the probe
oligonucleotides, with the result that a substrate-molecule complex is
formed, wherein the magnetic beads represent the substrate. As an
alternative, antibodies that bind specific target proteins or nucleic
acids can also be provided on the magnetic beads. The antibodies can be
polyclonal or monoclonal antibodies. As an alterative, at least one
device/element which bind nucleic acids non-specifically, e.g. silanes,
randomized oligonucleotides or the like, can also be provided on the
magnetic beads. Furthermore, it is conceivable for other substances that
bind specific biological molecules and structures, e.g. carbohydrates,
lipopolysaccharides and the like, to be applied on the beads.

[0081] In accordance with one alternative embodiment, the magnetic beads
can also be provided in the chamber 3 and have binding properties (e.g.
antibodies, polysaccharides, and the like) which specifically bind
specific biological structures in the sample, e.g. specific cells,
bacteria or viruses.

[0082] The process chambers are interconnected by microchannels 23, 25, 27
and 29 in accordance with the order of the process steps that proceed,
said microchannels being embodied in such a way that an interfering
exchange of liquid between the process chambers is largely prevented
during the processing and analysis and has no interfering influence. On
the other hand, the microchannels 23, 25, 27 and 29 are large enough to
permit magnetic beads 21 with bound structures or molecules to pass
through. The diameter of the microchannels 23, 25, 27 and 29 is typically
of the order of magnitude of several μm. As an alternative, with
larger dimensioning of the microchannels, it is also possible to provide
valves in the microchannels 23, 25, 27, 29 in order to fluidically
separate the individual process chambers 3, 5, 7, 9, 11 from one another
during the method sequence. In the chamber 5, the disruption of the
biological structures can be completed and non-bound sample constituents
can be separated from the molecules bound to the substrate (that is to
say the magnetic beads) by subsequent rinsing with washing solution or
buffer.

[0083] A further washing chamber 7 is provided in order to eliminate cell
residues and other contaminants that are possibly still present. The
complexes composed of magnetic beads 21 and nucleic acids (or composed of
magnetic beads and proteins in the case of a protein-binding property of
the magnetic beads) are moved through the microchannel 25 into the
washing chamber 7. By way of example chaotropic salts 35 can be stored in
the washing chamber 7, which salts are initially present in dry form and
dissolve as a result of the washing chamber 7 being filled.

[0084] For complex processing and analysis methods it is possible to
provide additional chambers.

[0085] The DNA molecules bound to the magnetic beads 21 are usually
present in a very low initial concentration in the sample, such that
amplification of the nucleic acids has to take place for detection. For
this purpose, the magnetic beads are moved into an amplification chamber
9, which is connected to the washing chamber 7 via the microchannel 27.
An amplification, for example by way of polymerase chain reaction (PCR)
or some other suitable amplification method, can take place in the
amplification chamber 9. The reagents 37 required for the amplification
reaction can be stored beforehand, e.g. in dry form, in the chamber 9.
The arrangement contains a peltier element, by way of which thermal
cycles can be carried out for the PCR reaction in the amplification
chamber 9. As an alternative, other heating and/or cooling elements known
to the person skilled in the art can also be present, e.g. a resistance
heating element or a water cooling system. The construction of the
arrangement is shown schematically in FIGS. 4 to 7, which will be
discussed in detail below.

[0086] When the temperature is increased, the DNA molecules are generally
detached from the magnetic beads 21. Consequently, the nucleic acids are
then released for an amplification reaction and a later detection
reaction. As an alternative it is also possible to amplify the nucleic
acids using the oligonucleotides applied on the beads as a primer for the
PCR reaction directly on the oligonucleotides. For this purpose, by way
of example, a corresponding primer for the counter-strand can
additionally also be provided in the amplification chamber, such that the
amplified nucleic acids are then bound to the magnetic beads at one end
via the probe oligonucleotides.

[0087] In order to detect the DNA, the nucleic acids bound to the magnetic
beads 21 can be moved through a microchannel 29 into a detection chamber
11. Specific oligonucleotides in a detection unit are immobilized in the
detection chamber 11. The amplified nucleic acids which are immobilized
on the magnetic beads at one end hybridize with the probe
oligonucleotides on the microarray and are thereby immobilized. The
detection of the nucleic acid molecules sought takes place by detection
of the immobilized magnetic beads at that location of the detection unit
39 at which the complementary oligonucleotides are arranged. For this
purpose, the detection unit 39 comprises a sensor that can detect the
presence of the magnetic beads 21 on the basis of the magnetic properties
thereof, e.g. a magnetoresistive sensor. As an alternative, it is
possible for the amplified nucleic acids hybridized to the probe
oligonucleotides of the microarray to be detected optically, e.g. by way
of fluorescent dyes, electrochemically, e.g. by redox cycling, or in some
other way.

[0088]FIG. 2 illustrates a further embodiment of a microfluidic device of
the arrangement according to an embodiment of the invention in the form
of a cartridge 1'. The cartridge 1' has four groups of process chambers
2, 4, 6, 8 respectively arranged along 4 reaction paths 10, 12, 14, 16.
Via corresponding filling openings 13, samples are introduced into the
cartridge and pass through the respective process chambers 2, 4, 6, 8
along the reaction paths 10, 12, 14, 16. It is noted at this point that
only the process chambers along the reaction path 10 are designated by
the reference symbols 2, 4, 6 and 8 in FIGS. 2 and 3, for reasons of
clarity; the corresponding process chambers along the reaction paths 12,
14, 16 should likewise be designated by these reference symbols. A
detection unit 39 of the type described above is provided in the
detection chamber 8. In this way, four samples can be processed an
analyzed in parallel in the cartridge 1'. It is also conceivable for two,
three, or 5 or more, e.g. 10 or 20 sample sections or reaction paths to
be arranged on a cartridge.

[0089] The expression "reaction path" denotes the path taken by the sample
or the biological molecules to be examined in the method sequence through
the device.

[0090]FIG. 3 shows a further alternative embodiment of a microfluidic
device in the form of a cartridge 1''. In this embodiment, four groups of
reaction chambers 2, 4, 6 are likewise arranged along four reaction paths
10, 12, 14, 16, such that four samples can be processed in parallel. The
samples are conducted along the reaction paths 10, 12, 14, 16 through the
respective process chambers 2, 4, 6 and are then conducted into a common
detection chamber 18, in which a common detection unit 39' is provided.

[0091] FIGS. 4 to 6 illustrate an arrangement 100 according to an
embodiment of the invention in different operating states, which
arrangement contains microfluidic devices 101, 101', 101'', 101a, 101b,
101c, 101d. A plurality of microfluidic devices 101 are stacked in a
magazine 103 embodied in stack-like fashion. The devices can already be
filled with samples before being introduced into the magazine 103. As a
result of an opening element 105 being opened and the transport units
107, 109 being advanced, the microfluidic device 101' is conveyed out of
the magazine 103. In the present example, in the arrangement 100 the
transport unit embodied as conveyor belts 107, 109 defines a central
transport section for the microfluidic devices 101, 101', which forms a
receptacle of the arrangement 100 for the microfluidic devices 101, 101'.
At least one device/element for fixing the magnetic beads 121 (e.g. in
the form of an electromagnet) and detection device 123 are provided along
this transport section. This operation is coordinated by the controller
111, 117, 119. A microfluidic device 101'' that had already been
processed previously has been transported into the collecting container
131.

[0092]FIG. 5 shows the arrangement according to an embodiment of the
invention in a further operating state, which temporarily succeeds the
operating state in accordance with FIG. 4. The microfluidic device 101'
is moved under a magnetic field generator 121 by the transport devices
107, 109. The magnetic field generator 121 can be embodied as a permanent
magnet or as an electromagnet. The process chambers 102, 104, 106, 108
provided in the device 101' embodied as a cartridge can be moved through
under the magnetic field generator 121 by the transport device 107, 109.
Through selective application of the magnetic field, the
substrate-molecule complex is fixed under the magnetic field generator,
while the microfluidic device 101' continues to move. As a result, the
molecules bound by the substrate are moved successively through the
process chambers 102, 104, 106, 108. As an alternative, however, it is
also possible to provide a moveable magnetic field generator which, with
an immobile cartridge, moves the sample bound to magnetic beads relative
to the cartridge. If an electromagnet is used, the magnetic field can be
controlled (e.g. on/off) by the controller 111. The microfluidic device
101' is moved further toward the right by the transport device 109. The
opening element 105 is then closed again.

[0093] After the microfluidic device 101' has been moved through under the
magnet 121, the detection chamber 108 is then situated under a sensor 123
(FIG. 6), which can read out the signals from the detection unit in the
detection chamber 108 in order to detect the presence or the
concentration of biological molecules to be examined, e.g. nucleic acids.
The detected signals can be conducted to the controller 111 and be
supplied there for data processing. After the signals have been detected,
the microfluidic device 101' can be transported into the collecting
container 131. The entire method sequence can then be repeated with the
next microfluidic device 101 situated in the magazine, until all the
samples have been processed. In this way, after the microfluidic devices
101 have been charged with the samples and the microfluidic devices 101
have been introduced into the magazine 103, it is possible for the entire
number of samples to be processed without necessitating further
intervention on the part of the operating personnel. Consequently, the
entire analysis of the samples can proceed in automated fashion.

[0094]FIG. 7 shows an alternative embodiment of the arrangement according
to the invention. Unfilled single-use microfluidic devices 101 configured
as a cartridge are supplied in rolled-up form in the magazine 103'. The
microfluidic devices can be rolled up e.g. on a flexible carrier strip.
In order to analyze the samples, firstly a microfluidic device 101a is
unrolled from the drum 104 and transported by the transport unit 107 to a
unit for introducing the samples 113.

[0095] The unit can be configured e.g. in the form of a moveable pipetting
arm. The samples are introduced into the microfluidic device 101a. The
entire method proceeds like an assembly line; while the samples are
introduced into the microfluidic device 101a, the microfluidic device
101b is moved under the magnet 121, with the result that the lysis and
washing steps are carried out in the corresponding process chambers. The
microfluidic device 101c is already situated under the sensor 123, where
the signals are read out from the detection unit in the microfluidic
device 101c. The microfluidic device 101d is transported into the
collecting container 131, which already contains a used microfluidic
device 101e.

[0096] In the manner illustrated a high number of samples can be processed
in automated fashion, the risk of contaminations or operating errors
being minimized. In particular by using cartridges on which a plurality
of samples can be processed in parallel, a high sample throughput can be
achieved in this way.

[0097] It is emphasized that the examples used are merely by way of
example and illustrative. Many different variations are conceivable in
particular with regard to the arrangement of components, the direction of
the movement of the cartridge through the arrangement, which could also
be circular, for example, and the sequence of lines and process chambers
in the cartridge.

[0098] Example embodiments being thus described, it will be obvious that
the same may be varied in many ways. Such variations are not to be
regarded as a departure from the spirit and scope of the present
invention, and all such modifications as would be obvious to one skilled
in the art are intended to be included within the scope of the following
claims.